Abstract
A proteinase of Mr 27,000 with a possible role in the metabolism and invasion of host tissues was purified from the conditioned medium of Trichophyton rubrum by concanavalin A and anion-exchange chromatography. Peaks of proteolytic activity were analyzed by substrate gel electrophoresis. The 27,000-Mr proteinase had a pH optimum of 8.0, a calcium dependence of 2 mM, and was inhibited by serine proteinase inhibitors, especially phenylmethylsulfonyl fluoride and Phe-Gly-Ala-Leu-chloromethyl ketone. By polyacrylamide gel electrophoresis, the 27,000-Mr proteinase had a reduced molecular weight of 44,000 and reacted with [3H]diisopropyl fluorophosphate. The proteinase degraded azocoll, type III collagen, type IV procollagen, laminin, fibronectin, and the peptide substrates succinyl-Ala-Ala-Pro-Phe-p-nitroanilide (1,573 M-1 s-1) and t-butyloxy carbonyl-Ala-Ala-Leu-p-nitroanilide (1,614 M-1 s-1).
Full text
PDF![3072](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c035/260772/c6ae66ffa96e/iai00070-0156.png)
![3073](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c035/260772/eb19ed35b3e4/iai00070-0157.png)
![3074](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c035/260772/9b2fe5d41890/iai00070-0158.png)
![3075](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c035/260772/d01a70b0e724/iai00070-0159.png)
![3076](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c035/260772/408dacf7f13c/iai00070-0160.png)
![3077](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c035/260772/c2ad6aa000c9/iai00070-0161.png)
![3078](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c035/260772/b394bd7f6089/iai00070-0162.png)
![3079](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c035/260772/9991e2d91282/iai00070-0163.png)
![3080](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c035/260772/3f54a8defd8d/iai00070-0164.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Allore R. J., Barber B. H. A recommendation for visualizing disulfide bonding by one-dimensional sodium dodecyl sulfate--polyacrylamide gel electrophoresis. Anal Biochem. 1984 Mar;137(2):523–527. doi: 10.1016/0003-2697(84)90121-0. [DOI] [PubMed] [Google Scholar]
- Apodaca G., McKerrow J. H. Regulation of Trichophyton rubrum proteolytic activity. Infect Immun. 1989 Oct;57(10):3081–3090. doi: 10.1128/iai.57.10.3081-3090.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Asahi M., Lindquist R., Fukuyama K., Apodaca G., Epstein W. L., McKerrow J. H. Purification and characterization of major extracellular proteinases from Trichophyton rubrum. Biochem J. 1985 Nov 15;232(1):139–144. doi: 10.1042/bj2320139. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- ERLANGER B. F., KOKOWSKY N., COHEN W. The preparation and properties of two new chromogenic substrates of trypsin. Arch Biochem Biophys. 1961 Nov;95:271–278. doi: 10.1016/0003-9861(61)90145-x. [DOI] [PubMed] [Google Scholar]
- Jones P. A., DeClerck Y. A. Destruction of extracellular matrices containing glycoproteins, elastin, and collagen by metastatic human tumor cells. Cancer Res. 1980 Sep;40(9):3222–3227. [PubMed] [Google Scholar]
- Laemmli U. K., Favre M. Maturation of the head of bacteriophage T4. I. DNA packaging events. J Mol Biol. 1973 Nov 15;80(4):575–599. doi: 10.1016/0022-2836(73)90198-8. [DOI] [PubMed] [Google Scholar]
- Lynch M. H., O'Guin W. M., Hardy C., Mak L., Sun T. T. Acidic and basic hair/nail ("hard") keratins: their colocalization in upper cortical and cuticle cells of the human hair follicle and their relationship to "soft" keratins. J Cell Biol. 1986 Dec;103(6 Pt 2):2593–2606. doi: 10.1083/jcb.103.6.2593. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McKerrow J. H., Jones P., Sage H., Pino-Heiss S. Proteinases from invasive larvae of the trematode parasite Schistosoma mansoni degrade connective-tissue and basement-membrane macromolecules. Biochem J. 1985 Oct 1;231(1):47–51. doi: 10.1042/bj2310047. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McKerrow J. H., Pino-Heiss S., Lindquist R., Werb Z. Purification and characterization of an elastinolytic proteinase secreted by cercariae of Schistosoma mansoni. J Biol Chem. 1985 Mar 25;260(6):3703–3707. [PubMed] [Google Scholar]
- Meevootisom V., Niederpruem D. J. Control of exocellular proteases in dermatophytes and especially Trichophyton rubrum. Sabouraudia. 1979 Jun;17(2):91–106. doi: 10.1080/00362177985380141. [DOI] [PubMed] [Google Scholar]
- Moll R., Franke W. W., Schiller D. L., Geiger B., Krepler R. The catalog of human cytokeratins: patterns of expression in normal epithelia, tumors and cultured cells. Cell. 1982 Nov;31(1):11–24. doi: 10.1016/0092-8674(82)90400-7. [DOI] [PubMed] [Google Scholar]
- NOVAL J. J., NICKERSON W. J. Decomposition of native keratin by Streptomyces fradiae. J Bacteriol. 1959 Mar;77(3):251–263. doi: 10.1128/jb.77.3.251-263.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nagle R. B. Intermediate filaments: a review of the basic biology. Am J Surg Pathol. 1988;12 (Suppl 1):4–16. [PubMed] [Google Scholar]
- RAUBITSCHEK F. Mechanical versus chemical keratolysis by dermatophytes. Sabouraudia. 1961 Jun;1:87–90. doi: 10.1080/00362176285190191. [DOI] [PubMed] [Google Scholar]
- Sanyal A. K., Das S. K., Banerjee A. B. Purification and partial characterization of an exocellular proteinase from Trichophyton rubrum. Sabouraudia. 1985 Jun;23(3):165–178. doi: 10.1080/00362178585380271. [DOI] [PubMed] [Google Scholar]
- Sun T. T., Green H. Keratin filaments of cultured human epidermal cells. Formation of intermolecular disulfide bonds during terminal differentiation. J Biol Chem. 1978 Mar 25;253(6):2053–2060. [PubMed] [Google Scholar]
- Takiuchi I., Higuchi D., Sei Y., Koga M. Immunological studies of an extra-cellular keratinase. J Dermatol. 1983 Aug;10(4):327–330. doi: 10.1111/j.1346-8138.1983.tb01146.x. [DOI] [PubMed] [Google Scholar]
- Takiuchi I., Higuchi D., Sei Y., Koga M. Isolation of an extracellular proteinase (keratinase) from Microsporum canis. Sabouraudia. 1982 Dec;20(4):281–288. [PubMed] [Google Scholar]
- Takiuchi I., Sei Y., Takagi H., Negi M. Partial characterization of the extracellular keratinase from Microsporum canis. Sabouraudia. 1984;22(3):219–224. [PubMed] [Google Scholar]
- Tsuboi R., Ko I. J., Matsuda K., Ogawa H. A new keratinolytic proteinase from clinical isolates of Trichophyton mentagrophytes. J Dermatol. 1987 Oct;14(5):506–508. doi: 10.1111/j.1346-8138.1987.tb03616.x. [DOI] [PubMed] [Google Scholar]
- WEARY P. E., CANBY C. M., CAWLEY E. P. KERATINOLYTIC ACTIVITY OF MICROSPORUM CANIS AND MICROSPORUM GYPSEUM. J Invest Dermatol. 1965 May;44:300–310. doi: 10.1038/jid.1965.54. [DOI] [PubMed] [Google Scholar]
- Wawrzkiewicz K., Lobarzewski J., Wolski T. Intracellular keratinase of Trichophyton gallinae. J Med Vet Mycol. 1987 Aug;25(4):261–268. [PubMed] [Google Scholar]
- Weary P. E., Canby C. M. Further observations on the keratinolytic acitivity of Trichophyton schoenleini and Trichophyton rubrum. J Invest Dermatol. 1969 Jul;53(1):58–63. doi: 10.1038/jid.1969.108. [DOI] [PubMed] [Google Scholar]
- Weary P. E., Canby C. M. Keratinolytic activity of trichophyton schoenleini , Trichophyton rubrum and trichophyton mentagrophytes. J Invest Dermatol. 1967 Mar;48(3):240–248. doi: 10.1038/jid.1967.38. [DOI] [PubMed] [Google Scholar]
- Yu R. J., Harmon S. R., Blank F. Hair digestion by a keratinase of Trichophyton mentagrophytes. J Invest Dermatol. 1969 Aug;53(2):166–171. [PubMed] [Google Scholar]
- Yu R. J., Harmon S. R., Blank F. Isolation and purification of an extracellular keratinase of Trichophyton mentagrophytes. J Bacteriol. 1968 Oct;96(4):1435–1436. doi: 10.1128/jb.96.4.1435-1436.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Yu R. J., Harmon S. R., Grappel S. F., Blank F. Two cell-bound keratinases of Trichophyton mentagrophytes. J Invest Dermatol. 1971 Jan;56(1):27–32. doi: 10.1111/1523-1747.ep12291869. [DOI] [PubMed] [Google Scholar]
- Yu R. J., Harmon S. R., Wachter P. E., Blank F. Amino acid composition and specificity of a keratinase of Trichophyton mentagrophytes. Arch Biochem Biophys. 1969 Dec;135(1):363–370. doi: 10.1016/0003-9861(69)90551-7. [DOI] [PubMed] [Google Scholar]